KR20190133482A - An anti-obesity or body fat reducing composition containing extract from a white kidney bean and a mushroom - Google Patents

Abstract

Disclosed is a body fat reduction and anti-obesity composition containing a white kidney bean extract and a Pleurotus eryngii extract as active components. The present invention provides a use of anti-obesity or body fat reduction by mixing the white kidney bean extract and the Pleurotus eryngii extract. A composition and pharmaceutical or functional food containing the same of the present invention have an excellent anti-obesity or body fat reduction effect.

Description

Anti-obesity or body fat reducing composition containing extract from a white kidney bean and a mushroom}

The present invention relates to a composition for reducing obesity or body fat, and more particularly, to a composition for preventing, treating and improving obesity comprising white kidney bean extract and agar mushroom extract as an active ingredient.

The composition for preventing and treating obesity comprising the white kidney bean extract and the extract of Awi mushrooms may be used as a pharmaceutical composition and / or health functional food composition to include a pharmaceutically acceptable excipient and / or a food acceptable acceptable excipient. Can be used.

The World Health Organization (WHO) has determined that obesity exceeds 30 kg / m ² of body mass index, with abnormal or excessive fat accumulating in the adipose tissue to the point of harm to health. The Asia-Pacific countries, including Korea, are over 25kg / m ² for obesity and 30kg / m ² for high obesity.

Obesity is a disease caused by a combination of factors such as environmental, genetic, mental and endocrine factors. Coronary artery disease, hypertension, stroke, type 2 diabetes, dyslipidemia, Sleep apnea, lung disease, and cancer. Obesity is a high socioeconomic cost because it is directly involved in the increased mortality from these diseases.

The prevalence of obesity in the past two decades has risen markedly and continues to rise globally, and in Korea, according to the 2015 National Health and Nutrition Survey, 33.2% (male: 39.7%, female 26.5%) of the adult population were surveyed as obesity. One out of three adults was found to be obese.

According to a white paper published by the National Health Insurance Corporation in 2016, the prevalence of obesity has increased rapidly from 29.3% in 2002 to 31.7% in 2013. According to the report of the National Institute of Health Insurance, Korea's Health Insurance Policy Research Institute, the socioeconomic cost of obesity in 2013 was 6.77 trillion won, an increase of 2.22 times compared to 3.5 trillion won in 2005. In contrast, about 17 times increase. Considering that the socioeconomic cost of smoking increased by 1.62 times and the socioeconomic cost of drinking increased by 1.56 times compared with 2005, the loss of obesity has recently increased. Medical expenses related to obesity including overweight rose 2.6 times from 1.7 trillion won in 2005 to 4.4 trillion won in 2013.

Currently, the anti-obesity agents sold in Korea include Belvic (component Lorcaserin), Contrab (component Bupropion / Naltrexone), purimine (component Phentemine), Puring (component Phendimetrazine), and saccharin (Glucagon-Like Peptide 1). The ultimate treatment goal of obesity is to go beyond weight loss and to ameliorate and prevent the diseases associated with obesity. Therefore, anti-obesity drugs should be safer in 'long-term use' and have an advantage in managing various comorbid diseases. However, there are a number of side effects that are currently on the market, including dizziness, nausea, constipation, dry mouth and depression.

A total of 27 raw materials are sold on the market for functional foods related to body fat reduction. Representative raw materials include Garcinia cambogia extract, complex extracts such as seaweed and wild mango seed extract. Garcinia cambogia extract has undergone functional re-evaluation from the Ministry of Food and Drug Safety due to safety issues related to liver disease, and complex extracts such as seaweed are also lacking in human test results to prove their effectiveness.

Most of the products that are sold as health functional foods for reducing body fat in Korea are imported and sold from overseas. Therefore, there is an urgent need to develop new raw materials for reducing body fat, which have been scientifically proven to be safe and effective during long-term use.

Korean Unexamined Patent Publication No. 2009-0007651

The present invention is to provide a composition having little side effects and safe for the human body, and excellent in preventing obesity or reducing body fat, and a pharmaceutical or functional food containing the same.

In order to achieve the above technical problem, the present invention provides an anti-obesity or body fat reduction composition containing the extract of white kidney beans and awi mushrooms as an active ingredient used as food ingredients.

The present invention also provides an anti-obesity or body fat-reducing medicine or health functional food comprising the anti-obesity or body fat-reducing composition.

The present invention provides a use of anti-obesity or body fat reduction by mixing a white kidney bean extract and a mushroom extract. The composition of the present invention and a pharmaceutical or functional food containing the same have an excellent anti-obesity or body fat reduction effect.

1 is a process chart showing a process for preparing a composition according to the present invention.
Figure 2 is a graph showing the α-amylase inhibition rate by mixing ratio of the anti-obesity composition according to the present invention using the α-amylase (amylase) inhibition test.
Figure 3 is a graph showing the lipase activity rate of the mixing ratio of the anti-obesity composition according to the present invention using a triglyceride lipase activity test.
Figure 4 is a graph showing the change in body fat according to the mixing ratio of the anti-obesity composition in a high fat diet-induced obesity model.
Figure 5 is a graph showing the weight change according to the mixing ratio of the anti-obesity composition in a high fat diet-induced obesity model.
Figure 6 is a graph showing the change in weight measurement according to the dose of the anti-obesity composition after 8 weeks repeated administration in a high fat diet-induced obesity model by date and experimental groups, respectively.
Figure 7 is a graph showing the total fat, abdominal fat and subcutaneous fat change with respect to the change in body fat mass according to the dose of the anti-obesity composition after repeated administration for 8 weeks in a high-fat diet obesity model.
8 is a graph showing the change in carbohydrate content in the feces according to the dose of the anti-obesity composition after 8 weeks repeated administration in a high fat diet-induced obesity model.
9 is a graph showing triglyceride changes in the liver according to the dose of the anti-obesity composition after repeated 8-week administration in a high fat diet-induced obesity model.

The present inventors endeavored to obtain a composition having anti-obesity and body fat reduction effect among natural products which have been used and consumed for a long time, and confirmed that the mixed extract of white kidney beans and awi mushrooms has an excellent anti-obesity and body fat reduction effect. The present invention has been completed.

Therefore, the present invention relates to a composition for preventing, treating or improving obesity, including white kidney bean extract and agar mushroom extract as active ingredients. In addition, the present invention relates to a composition for preventing, treating or improving obesity comprising an extract of a mixture of white kidney beans and awi mushroom as an active ingredient.

The anti-obesity or body fat reducing composition of the present invention can be obtained through conventional methods such as extraction, concentration and drying. For example, in the present invention, the extract may be obtained by extracting with water, alcohol (preferably 1 to 4 carbon straight chain or branched alcohol), or a mixture of water and alcohol. In addition, the extract thus obtained may be obtained by further extraction with a nonpolar solvent such as hexane, dichloromethane, chloroform, or ethyl acetate. In the present invention, the extract may be extracted by applying heat during extraction. Preferably, the extract is extracted by applying heat when extracting the extract.

In the present invention, the content ratio of the white kidney bean extract and the awi mushroom extract is 1: 1 to 1: 5 and 1: 1 to 5: 1, preferably 1: 1 to 5, based on the weight of the solid dried extract. : 1, more preferably 2: 1 to 4: 1.

The anti-obesity or body fat reducing composition of the present invention can be prepared in the form of pharmaceuticals and functional foods. Pharmaceuticals and health functional foods may include pharmaceutically or pharmaceutically acceptable excipients or additives.

The pharmaceutical or functional food containing the anti-obesity or body fat reducing composition of the present invention may be a solid preparation or a liquid preparation. Solid form preparations include, but are not limited to, tablets, capsules, granules, powders, transdermals, suppositories, and the like. Solid form preparations may include excipients, binders, disintegrants, glidants, coatings and the like. Liquid formulations include, but are not limited to, aqueous solutions, suspensions, emulsions, syrups, aerosols, or sterile injectable solutions, and can be prepared by adding suitable colorants, flavors, stabilizers, and the like.

For example, tablets may be mixed with suitable pharmaceutically acceptable excipients (maltodextrin) and pharmaceutically acceptable suitable binders such as D-sorbitol powder, silicon dioxide, and pharmaceutically acceptable suitable glidants such as magnesium stearate. It can be prepared using a tablet press.

The anti-obesity or body fat reducing composition of the present invention may be administered as an oral, injectable, inhalant, etc., depending on the condition to be treated and the condition of the individual, but is not limited thereto. It may be formulated into a suitable formulation including non-toxic, pharmaceutically acceptable carriers, additives, and the like, commonly used according to the route of administration.

Oral liquid preparations include suspensions, solvents, emulsions, and syrups.In addition to the commonly used simple diluents, water and liquid paraffin, various excipients may be included, such as wetting agents, sweeteners, fragrances, and preservatives. have. Formulations for parenteral administration include sterile aqueous solutions, suspensions, emulsions, and lyophilized preparations.

In order to achieve the object of the invention, such as anti-obesity or body fat reduction, it is preferable that the composition has a daily dose of 1.5 g / day or less of an active ingredient based on animal test results. For example, the composition may be administered in an amount of 0.1 to 300mg / kg daily dose based on the active ingredient. The dosage may vary depending on the patient's condition (age, weight, gender, etc.). If necessary, the total daily dose may be divided into one day for convenience.

Hereinafter, examples and the like will be described in detail to help understand the present invention. However, embodiments according to the present invention may be modified in many different forms, and the scope of the present invention should not be construed as being limited by the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

[ Example  One] White Kidney Beans  And From a mushroom  Preparation of the composition according to the invention

White kidney beans ( Phalotus multiflorus var. Albus Bailey ) and agaric mushroom ( Pleurotus ) according to the process as shown in Figure 1 ferulae ) extract was prepared.

Specifically, the extract-containing composition was obtained as follows.

Purified water was added 3 to 10 times to 5 kg of white kidney beans dry powder, and extracted with stirring for 24 hours. The extracted extract was filtered (No. 2 whatman, Maidstone, England) and the filtrate was concentrated under reduced pressure (rotary vacuum evaporator) to obtain an extract. The extract was freeze-dried (PVTFD10R, ILSHIN, Daejeon, Korea) to finally obtain 1.07 kg white kidney bean extract powder.

Purified water was added 3 to 10-fold to 5 kg of the above-mentioned mushroom dry powder, and extracted with stirring at 80 ° C for 10 hours. The prepared extract was filtered and the filtrate was concentrated under reduced pressure to obtain an extract. Dextrin was added to the extract, followed by spray drying. Finally, 1.01 kg of Awi mushroom extract powder was obtained.

White kidney bean extract powder and awi mushroom extract powder were stored in a freezer at -20 ° C., respectively, and the extracts were used alone or in the following experiments.

[ Example  2] α-amylase inhibition test Anti-obesity  Effect evaluation

α-amylase inhibitory activity was evaluated by measuring the amount of soluble starch decreased by the enzymatic reaction by the iodine reaction method.

Human saliva-derived α-amylase was dissolved in PBS (phosphate buffer) at a concentration of 20 units / ml, and soluble starch was used as a substrate of α-amylase in 1% concentration in PBS (phosphate buffer).

To measure the inhibitory activity against α-amylase, 290 μl PBS, 10 μl α-amylase solution (20 units / ml) and 50 μl extract (1.0 mg / ml) were mixed and incubated at 37 ° C. for 10 minutes.

To the incubated mixture, 350 μl of 1% soluble starch as a substrate was added, and again incubated at 37 ° C. for 30 minutes to react.

To measure the amount of soluble starch remaining after the reaction, 300 μl of iodine solution (0.1% KI + 0.01% I ₂ /0.05N HCl) was added to the reaction solution (700 μl), and the absorbance was measured at 620 nm using a spectrophotometer. Measured.

As a standard of the α-amylase inhibitor, it was dissolved in DMSO at a concentration of 100 μg / ml of acarbose, and compared with the extract.

The test group was evaluated with white kidney bean extract powder, awi mushroom extract powder, mixed extract powder (1: 1 ~ 1: 5 and 1: 1 ~ 5: 1) group of white kidney bean and awi mushroom (concentration of mixed extract) Maintain 1.0 mg / ml). In the control group, acarbose was used.

Table 1 below summarizes the composition and test procedure of the control group, test group, black 1 and 2.

Composition of the Test Composition Control Extract test group Blank 1 Blank 2 α-amylase solution (20 units / ml) 10 μl 10 μl PBS buffer 290 μl 290 μl 300 μl 300 μl Inhibitors (White Kidney Bean and Agaric Mushroom Extract) - 50 μl - 50 μl Standard (Akaboz) DMSO Solution 50 μl - 50 μl - Free incubation (37 ℃, 10 minutes) 1% Soluble Starch Solution 350 μl 350 μl 350 μl 350 μl Incubation (37 ℃, 10 minutes) I ₂ solution (0.1% KI + 0.01% I ₂ /0.05N HCl) 300 μl 300 μl 300 μl 300 μl OD measurement (620nm)

The measurement results are shown in FIG. As shown in Figure 2, the anti-obesity composition according to the present invention increased the α-amylase inhibitory activity as the ratio of white kidney beans extract powder increases.

 [ Example  3] TGL ( triglyceride  lipase activity test Anti-obesity  Effect evaluation

Lipase is responsible for breaking down triglycerides. Orlistat, a typical obesity treatment, reduces the absorption of triglycerides and cholesterol in the body by irreversibly binding and inactivating lipases that degrade triglycerides. Triglycerides and cholesterol that are not degraded are excreted unchanged in feces.

Orlistat as a standard for the evaluation of triglyceride lipase (TGL) activity was dissolved in DMSO at a concentration of 100 μg / ml and compared with the extract.

16 μl substrate mixture (5 μl of 120 mM Tauricholic acid solution + 1 M Enzyme Assay buffer (1 M NaH ₂ PO ₄ , 1 M NaCl, pH 7.0) 10 ul + 400 mM P-NPB (p-nitrophenyl butyrate) 1 ul) was added to each well (96well). plate). In each well, add 54 ul of standard orlistat-containing DMSO solution or 54 ul of extract (concentration 250 μg / ml). Absorbance is measured at 405 nm using a plate reader.

Enzyme (2 mg / ml TGL solution 10ul + 0.1% BSA buffer 20ul) was added to the wells and mixed, the light is blocked, and incubated at room temperature for 10 minutes. Absorbance is measured at 405 nm using a plate reader.

The test group was evaluated using a group of white kidney bean extract powder, awi mushroom extract powder, a mixture of white kidney bean extract and awi mushroom extract (1: 1 to 1: 5 and 1: 1 to 5: 1) (single extract and The mixed extract concentrations should all be 250 μg / ml). Negative control groups do not contain extracts or test substances.

The results are shown in FIG. As shown in Figure 3, the anti-obesity composition according to the present invention increased the activity of the triglyceride enzyme as the ratio of agaric mushrooms increased.

[ Example  4] High Fat Diet (45% HFD Evaluation of Anti-obesity Effect by Mixing Ratio in Extracts

Thirty-three C57BL / 6J male mice, 8 weeks old, were acclimated for a week, followed by feeding a high fat diet (45% HFD (Research diet, D12451) pellet form-free diet) for 2 weeks to induce obesity.

Two weeks later, the experiment was divided into five groups as follows:

(i) Negative control group that consumed only high fat diet [Con (-), n = 10], hereinafter referred to as 'Con (-)',

(ii) Positive control group (HFD + orlistat + acarbose, n = 8, hereinafter referred to as 'Con (+)') mixed with orlistat and acarbose (orlistat + acarbose) on a high-fat diet,

(iii) Experimental group 1 (HFD + mixture, n = 8, hereinafter referred to as 'Case1') to which the anti-obesity composition of the present invention (white kidney bean: awi mushroom 1: 1) was administered to a high fat diet,

(iv) Experimental group 2 (HFD + mixture, n = 8, hereinafter referred to as 'Case2') in which the anti-obesity composition of the present invention (white kidney bean: Awi mushroom 2: 1) was administered to a high fat diet,

(v) Experimental group 3 (HFD + mixture, n = 8, hereinafter referred to as 'Case3') in which the anti-obesity composition of the present invention (white kidney bean: Awi mushroom 3: 1) was administered to a high fat diet.

Negative and diet were freely fed and body fat mass and body weight were measured during the experiment. The test dose was administered at 200 mg / kg.

The anti-obesity composition [Case 3] mixed extract, the tendency to reduce the body fat mass and weight at 8 weeks compared to the high-fat diet group [Con (-)] was confirmed in Figures 4 and 5.

This means that ingestion of high fat diet increased body fat mass and body weight, and ingestion of the anti-obesity composition [Case3] of the present invention inhibited such body fat mass and weight gain.

[ Example  5] Different concentrations of extracts in high fat diet obesity induction model Anti-obesity  Effect evaluation

Animal testing and diet

Eighty-eight male C57BL / 6J males were divided into seven test groups for eight weeks (56 days), and the diet was applied.

The test group was divided into seven groups. In other words

(i) Group that consumed only regular diet (n = 12), hereinafter referred to as 'G1',

(ii) High-fat diet only group (n = 12), hereinafter referred to as 'G2',

(iii) A group ingested with a high-fat diet mixed with Zenical (orlistat 40mg / kg / day) (HFD + orlistat, n = 12, hereinafter referred to as 'G3'),

(iv) Group ingested by mixing Garcinia Cambogia (200mg / kg / day) in a high fat diet (HFD + Garcinia Cambogia, n = 12, hereinafter referred to as 'G4'),

(v) In the high fat diet, the group administered the anti-obesity composition of the present invention (white kidney bean: Awi mushroom 3: 1 100 mg / kg / day) (HFD + mixture, n = 12, hereinafter referred to as 'G5'),

(vi) In the high fat diet, the group administered the anti-obesity composition of the present invention (white kidney bean: Awi mushroom 3: 1 200 mg / kg / day) (HFD + mixture, n = 12, hereinafter referred to as 'G6'),

(vii) The group administered the anti-obesity composition of the present invention (white kidney bean: Awi mushroom 3: 1 300 mg / kg / day) in a high fat diet (HFD + mixture, n = 12, hereinafter referred to as 'G7').

Negative and diet were freely administered during the trial period, including: ① body weight (measured at the start of anti-obesity composition administration, twice a week thereafter and on autopsy day), and ② body fat (eight weeks after initiation of anti-obesity composition administration (before autopsy) Animal fat mass measured by micro-CT (micro-CT)), ③ Carbohydrate content in feces (measured by ELISA analysis for 8 weeks after administration of anti-obesity composition), ④ TG (triglyceride) in liver tissue Changes were measured (analyzed by blood samples collected at 4, 6, 7, and 8 weeks after the start of the anti-obesity composition administration and at the time of autopsy).

The composition and dose settings of each test group are shown in Table 2 below:

Test Group Configuration and Dose Setting group obesity
Induction The number of animals
(Mari) Substance Dosage
(mg / kg / day) Dosage
(mL / kg / day) Number of doses G1 N 12 D.W - 10 2 times / day G2 Y 12 D.W - 10 2 times / day G3 Y 12 Orlistat 40 10 2 times / day G4 Y 12 Garcinia Cambogia 200 10 2 times / day G5 Y 12 Low dose anti-obesity composition 100 10 2 times / day G6 Y 12 Medium Dose Anti-Obesity Composition 200 10 2 times / day G7 Y 12 High dose anti-obesity composition 300 10 2 times / day

The general symptoms and health status were confirmed, and 84 animals having a weight gain of 10% or more compared to the normal diet were used for the test.

(One) Weight change measurement

Body weights were measured on the day of administration of the anti-obesity composition, afterwards twice a week and on an autopsy day.

Before the administration of the anti-obesity composition, the weight level of all the obesity induction groups (G2-G7) was significantly higher than the normal control group (G1) (p <0.001). In addition, the induced control group (G2) showed significant weight gain over the entire observation period compared to the normal control group (G1) (p <0.001). The results are shown in FIG.

This means that the intake of a high-fat diet increased the weight, and that the ingestion of the anti-obesity composition high dose group (G7) of the present invention inhibited such weight gain.

(2) Fat mass analysis using micro CT

Eight weeks after the start of anti-obesity composition administration (before necropsy) was taken by Micro-CT (vivaCT 8-, SCANCO Medical, Switzerland) to measure the body fat mass of all animals. The measurement site of body fat was analyzed by setting abdominal fat and subcutaneous fat which exist in the space from the 2nd lumbar starting point to the 5th lumbar end (L2-L5).

The induced control group (G2) showed a significant increase in total fat, abdominal fat and subcutaneous fat compared to the non-induced control. The results are shown in FIG. 7 (p <0.001).

This means that the intake of a high-fat diet increased the total body fat, abdominal fat and subcutaneous fat in the body, and that the intake of the anti-obesity composition high dose group (G7) of the present invention inhibited such an increase in body fat.

(3) Feces  Carbohydrate Content Measurement

Before starting the anti-obesity composition administration, feces were collected by each breeding box once a week. Collected feces were stored in a cryogenic freezer (about -70 ℃ or less) until analysis, and analyzed by ELISA analysis for samples 2 and 8 weeks after the anti-obesity composition administration.

Fecal analysis showed a significant increase in carbohydrates at week 8 in the high dose anti-obesity composition-administered group (G7) compared to the induced control group (G2). This is shown in FIG. 8 (p <0.001).

This means that administration of the high dose anti-obesity composition (G7) inhibited the absorption of carbohydrates and inhibited the accumulation of fat in the body.

(4) Soy sauce Triglycerides ( triglyceride Content measurement

On the day of necropsy, animals were inhaled with anesthesia, and when anesthesia was confirmed, they were opened, and blood was collected using a syringe from the posterior vena cava. After removing the liver tissue, it was stored in an ultra-cold freezer (about -70 ℃ or less) until analysis, and analyzed by ELISA analysis.

As a result of analysis of TG (triglyceride) content in the liver, the induced control group (G2) showed a significant increase compared to the non-induced control group (G1) (p <0.001), and the high dose anti-obesity composition group (G7) compared to the induced control group (G2). Showed a significant decrease. This is shown in FIG. 9 (p <0.005).

This means that administration of the anti-obesity composition inhibited fat accumulation in the body by significantly reducing the elevated TG content by high fat diet.

The composition of the present invention is to provide a composition having a low side effect and safe for the human body, excellent in preventing obesity or reducing body fat, and a pharmaceutical or functional food containing the same.

Claims (14)

A composition for preventing, treating or improving obesity comprising white kidney bean extract and agaric mushroom extract. The composition of claim 1, wherein the white kidney bean extract is extracted using water, alcohol or a mixture of water and alcohol as an extraction solvent. The composition of claim 1, wherein the extract of Awi mushroom is extracted using water, alcohol or a mixture of water and alcohol as an extraction solvent. The composition of claim 1, wherein the white kidney bean extract and the agar mushroom extract are extracted with water, alcohol or a mixture of water and alcohol as an extractant. The composition according to any one of claims 1 to 4, wherein the awi mushroom extract is extracted by applying heat during extraction. The extract of any one of claims 2 to 4, wherein the white kidney bean extract or the above-mentioned mushroom extract is extracted with water, alcohol or a mixture of water and alcohol as an extraction solvent, and then further extracted with a nonpolar solvent as an extraction solvent. Composition. The composition according to any one of claims 1 to 4, wherein the content ratio of the white kidney bean extract and the awi mushroom extract is 1: 1 to 5: 1 based on the weight of the dried solids. The composition according to any one of claims 1 to 4, wherein the content ratio of the white kidney beans extract and the extract of Awi mushrooms is 2: 1 to 4: 1 based on the weight of the dried solids. A composition for preventing, treating or improving obesity comprising an extract of a mixture of white kidney beans and agar mushroom. The composition of claim 9, wherein the extract is extracted using water, alcohol or a mixture of water and alcohol as an extraction solvent. The composition of claim 10, wherein the extract is extracted by applying heat during extraction. The composition of claim 10, wherein the extract is extracted using water, alcohol or a mixture of water and alcohol as an extraction solvent, and then further extracted using a nonpolar solvent as the extraction solvent. A pharmaceutical composition or health functional food comprising the composition according to any one of claims 1 to 4 and 9 to 12. A pharmaceutical composition or health functional food comprising the composition of claim 7.

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